Copurification of the Lac repressor with polyhistidine-tagged proteins in immobilized metal affinity chromatography.

One of the commonly used resins for immobilized metal affinity purification of polyhistidine-tagged recombinant proteins is TALON resin, a cobalt (II)--carboxymethylaspartate-based matrix linked to Sepharose CL-6B. Here, we show that TALON resin efficiently purifies the native form of Lac repressor, which represents the major contaminant when (His)(6)-tagged proteins are isolated from Escherichia coli host cells carrying the lacI(q) gene. Inspection of the crystal structure of the repressor suggests that three His residues (residues 163, 173, and 202) in each subunit of the tetramer are optimally spaced on an exposed face of the protein to allow interaction with Co(II). In addition to establishing a more efficient procedure for purification of the Lac repressor, these studies indicate that non-lacI(q)-based expression systems yield significantly purer preparations of recombinant polyhistidine-tagged proteins.

Transparent electrode with a nanostructured coating.

Using single-walled nanotubes as an example, we fabricated transparent conductive coatings and demonstrated a new technique of centrifuge coating as a potential low-waste, solution-based batch process for the fabrication of nanostructured coatings. A theoretical model is developed to account for the sheet resistance exhibited by layered random-network coatings such as nanofilaments and graphene. The model equation is analytical and compact, and allows the correlation of very different scaling regimes reported in the literature to the underlying coating microstructure. Finally, we also show a refined experimental setup to systematically measure the curvature-dependent sheet resistance.